CN115634234B - Poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid and preparation and application thereof - Google Patents
Poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid and preparation and application thereof Download PDFInfo
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- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 title claims abstract description 79
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Medicinal Preparation (AREA)
Abstract
The invention relates to the technical field of medical products, in particular to poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid, and preparation and application thereof. Comprises hyaluronic acid, chlorhexidine, geniposide, poloxamer 407 and NaCl. The invention combines chlorhexidine, hyaluronic acid, naCl, geniposide and poloxamer 407, and successfully designs a temperature-sensitive gel for treating and repairing skin and mucous membrane wound surfaces through screening and optimizing prescriptions and processes, and the use of the hyaluronic acid ensures that the gel has excellent effects of absorbing, infiltrating, moisturizing and promoting wound surface healing. The invention can be used for spraying and administering large-area wounds, and an in-situ gel film can be formed within 30 seconds, so that patients can conveniently and automatically administer the medicine. For small-sized, concealed, slit and fold wounds, the administration can be by push injection, the good fluidity is convenient for infiltration, and the good gelatinization is convenient for adhesion and filling.
Description
Technical Field
The invention relates to the technical field of medical products, in particular to poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid, and preparation and application thereof.
Background
The skin and mucous membrane of the human body are barriers for maintaining stable environment and preventing invasion of microorganisms, and a series of problems of the human body, such as bacterial infection, aggravation of metabolism, excessive loss of water and protein, endocrine and immune system dysfunction, etc. are caused by skin and mucous membrane injury caused by ulcer, wound burn, bedsore, inflammation, etc., and the life may be seriously endangered.
There are dressing, ointment and gel for repairing skin and mucous membrane injury on the market. The dressing is fixed for a long time, but cannot reach the deeper region of the wound; the ointment is not suitable for mucous membrane, and has poor air permeability and strong discomfort for patients; the formed gel cannot penetrate into small gaps in the wound and is not easy to uniformly spread. The temperature-sensitive hydrogel has temperature-stimulated sol-gel transition behavior, is in a solution state at room temperature, can be conveniently applied to a lesion site in a pushing injection mode and the like, can permeate deep wounds and folds due to good fluidity, and then rapidly forms a gel film under the driving of body temperature, so that the effects of keeping the wounds moist, absorbing secretion, relieving pain and controlling bleeding are exerted, and the quick healing of the wounds is promoted.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a temperature-sensitive hydrogel for treating and repairing damaged skin and mucous membrane, which has the advantages of good adhesiveness, prolonged local effect of the medicament, short gelation time, proper viscosity and strength, good biocompatibility, simple preparation process, good patient compliance and antibacterial and anti-inflammatory effects, and preparation and application thereof. The temperature-sensitive hydrogel has strong antibacterial effect on gram-positive bacteria represented by escherichia coli and gram-negative bacteria represented by staphylococcus aureus, and has remarkable treatment and repair effects on mucosal ulcers. The thermosensitive hydrogel can be quickly attached to a wound surface to form an in-situ gel film after being applied, a physical protective barrier is provided for a damaged area, the medicine is ensured to stay and act on the damaged area only and is released at a constant rate, the curative effect is improved, the toxic and side effects of the whole body are reduced, and the medicine is convenient for a patient to administer by himself. The application of the 0.5% hyaluronic acid of the temperature-sensitive hydrogel can enhance protein accumulation in a wound surface, increase skin permeability and prevent bacterial invasion, and the effective moisture-preserving and air-permeable capacity of the temperature-sensitive hydrogel is beneficial to preventing wound infection, protecting the wound, and has the effects of inhibiting bacteria, resisting inflammation, stopping bleeding, easing pain and the like while promoting wound healing. The temperature sensitive hydrogel can be used for treating small-area skin, mucous membrane wound and ulcer, such as stomatocace, oral mucositis, etc., and large-area irregular wound, such as burn, impetigo, bedsore, tinea manus, tinea pedis, female colpitis, cervicitis, cervical erosion, etc.
Chitosan is the most commonly used temperature-sensitive gel matrix, but in order to make the chitosan have proper physical and chemical properties of gel, such as gelation temperature, gelation time, viscosity, strength, adhesive force and the like, chemical modification is needed, the process is complex, and the research and development cost is high. Poloxamer is a synthetic nonionic triblock copolymer (PEO-PPO-PEO), in which poloxamer 407 has good sol-gel transition behavior at body temperature. The literature shows that poloxamer 407 forms a gel at a concentration ranging from 15 to 30%. Our earlier studies found that the smaller the poloxamer 407 concentration, the lower the gel viscosity, mechanical strength formed. However, when the concentration of poloxamer 407 is high, on the one hand, no document proves that poloxamer 407 in high concentration has no cytotoxicity as low concentration; on the other hand, the formed gel is slow to degrade, and when the gel is used for treating mucosal ulcers, broken gel is not easy to remove. Therefore, some additives are added to adjust its properties for ideal application to skin and mucous membranes.
Hyaluronic acid is biocompatible, biodegradable and non-immunogenic, and can act as an anti-inflammatory agent by inhibiting cyclooxygenase to alter prostaglandin biosynthesis; can also play roles in relieving inflammation, helping healing after tissue injury and starting repair by regulating the recruitment of inflammatory cells, the release of inflammatory cytokines and the migration of cells. Several medical hyaluronic acid dressings are marketed in China. Because poloxamer 407 and hyaluronic acid can form intermolecular hydrogen bonds, the addition of hyaluronic acid can obviously improve the mechanical strength, viscosity and adhesive force of poloxamer 407. Therefore, the invention develops the thermosensitive hydrogel with antibacterial and anti-inflammatory effects by taking poloxamer 407-hyaluronic acid as a carrier.
Chlorhexidine is a biguanide preservative, is mature in clinical use and is listed as a bacteriostatic gold standard at home and abroad. It has broad bactericidal activity against gram positive and gram negative bacteria, and is effective against fungi and yeasts, including candida, and viruses such as hiv and hepatitis b.
The geniposide is a natural product extracted from dried mature fruits of Gardenia jasminoides Ellis, is an iridoid glycoside compound, and has good anti-inflammatory effect. Compared with widely used anti-inflammatory drugs such as glucocorticoid and antibiotics, the preparation has small toxic and side effects and high safety coefficient, and has obvious advantages in treating chronic inflammatory diseases.
Therefore, the invention selects chlorhexidine as a bacteriostatic agent and geniposide as an anti-inflammatory agent for researching the temperature-sensitive hydrogel. Experiments show that there is no incompatibility between the geniposide and the carrier, but a large amount of floccules are formed immediately after chlorhexidine is contacted with hyaluronic acid due to salt formation reaction. On the other hand, the poloxamer 407 modified by hyaluronic acid has high gel viscosity and strength and is not easy to adhere to the skin and mucous membrane. In order to overcome the problems, the invention performs a series of screening and optimizing of the preparation method and the additives, and successfully prepares the temperature-sensitive hydrogel with excellent antibacterial and anti-inflammatory activities, good adhesive force and proper physicochemical properties.
The invention is realized by the following technical scheme: the poloxamer 407 antibacterial anti-inflammatory thermosensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid comprises the following raw material components of hyaluronic acid, chlorhexidine, geniposide, poloxamer 407 and NaCl.
As a further improvement of the technical scheme of the temperature-sensitive hydrogel, the hyaluronic acid is 0.1-0.8%, chlorhexidine is 0.05-0.1%, geniposide is 0.5%, poloxamer 407 is 14.0-15.0% and NaCl is 0.05-0.2%.
As a further improvement of the technical scheme of the temperature-sensitive hydrogel, the hyaluronic acid is 0.5%, the chlorhexidine is 0.1%, the geniposide is 0.5%, the poloxamer 407 is 15.0% and the NaCl is 0.1-0.2%.
The invention further provides poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid, which comprises the following raw material components, by weight, 0.1-0.8% of hyaluronic acid, 0.05-0.1% of chlorhexidine, 0.5% of gardenin, 14.0-15.0% of poloxamer 407, 0.05-0.2% of NaCl, and the balance of water.
As a further improvement of the technical scheme of the temperature-sensitive hydrogel, the hyaluronic acid is 0.5%, the chlorhexidine is 0.1%, the geniposide is 0.5%, the poloxamer 407 is 15.0% and the NaCl is 0.1-0.2%.
The invention further provides a preparation method of the poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid, which comprises the following steps:
step 1): adding chlorhexidine into water for fully dissolving, adding poloxamer 407, standing, and standing at 4 ℃ overnight to fully swell and dissolve;
step 2): adding NaCl into water for dissolution, uniformly scattering hyaluronic acid on the surface of the NaCl, and standing the NaCl to enable the NaCl to be fully swelled and dissolved at room temperature;
step 3): mixing the solutions obtained in the step 1) and the step 2), uniformly stirring, adding the geniposide, and fully stirring and dissolving to obtain the temperature-sensitive hydrogel.
As a further improvement of the technical scheme of the preparation method, the chlorhexidine is one of chlorhexidine acetate, chlorhexidine gluconate and chlorhexidine hydrochloride.
The invention also provides a temperature-sensitive hydrogel prepared by the preparation method of the poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid.
The invention further provides application of the temperature-sensitive hydrogel in preparation of antibacterial anti-inflammatory drugs.
The invention further provides application of the temperature-sensitive hydrogel in preparing a medicament for treating and repairing skin and mucous membrane injury.
The beneficial effects of the invention are as follows:
1. the invention combines chlorhexidine, hyaluronic acid, naCl, geniposide and poloxamer 407, successfully designs a temperature-sensitive gel for skin and mucous membrane wound treatment and repair through screening and optimizing of prescriptions and processes, and the use of 0.5% hyaluronic acid ensures that the gel has excellent effects of absorbing, infiltrating, moisturizing and promoting wound healing.
2. The chlorhexidine is firstly coated in the poloxamer 407 and then mixed with the hyaluronic acid, so that the direct contact between the hyaluronic acid and the chlorhexidine is avoided, and the gold-plate bacteriostatic agent chlorhexidine and the hyaluronic acid which is an excellent skin mucosa repair material are successfully compatible.
3. After poloxamer 407 is modified by hyaluronic acid within the concentration range capable of forming temperature-sensitive gel, the viscosity and strength of the gel are obviously increased due to intermolecular hydrogen bonding, which is unfavorable for film formation and adhesion to wounds. The invention utilizes the principle that neutral electrolyte can shield the electroadhesion effect, adds a very small amount of NaCl to reduce the viscosity and strength of the gel, and screens the optimal concentration which can form the gel and is suitable for injection.
4. The precipitation occurs due to the salting-out effect when NaCl and chlorhexidine are simultaneously dissolved in the poloxamer 407 solution. According to the invention, naCl is dissolved in a hyaluronic acid aqueous solution through screening and then is mixed with a poloxamer 407 solution containing chlorhexidine, so that the temperature-sensitive hydrogel with proper appearance is successfully prepared, and the temperature-sensitive hydrogel has good stability. See fig. 1.
5. The acetic acid-induced wistar rat oral mucosa ulcer is taken as a model to evaluate the repairing effect of the product on the mucosa, and the obvious treatment effect is shown. As shown in fig. 4.
6. The invention has good inhibition effect on escherichia coli, staphylococcus aureus, candida albicans, pseudomonas aeruginosa, streptococcus pneumoniae and streptococcus pyogenes, and is obviously superior to the similar preparation of the commercial chlorhexidine. As shown in fig. 3.
7. The invention can be used for spraying and administering large-area wounds, can form an in-situ gel film within 30 seconds, and can be conveniently and automatically administered by patients, thereby avoiding medicine waste and discomfort of the patients, reducing systemic toxic and side effects and improving compliance of the patients.
8. For small, hidden, slit and fold wounds, such as alveoli, sublingual etc., the drug can be administered by bolus injection, with good fluidity for penetration and good gelling for attachment and filling.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a physical image of example 7 after 18 months of storage. In the figure, A is liquid at room temperature, and B is gel at body temperature. As can be seen from the figures: in the A graph, the temperature-sensitive hydrogel is in a liquid state in a room temperature environment, the liquid level is inclined along with the inclination of the test tube, the fluidity is good, and the administration in a spray or push injection mode is convenient; in the graph B, the temperature-sensitive hydrogel is changed into gel under the condition of body temperature, and when a test tube is inclined, the object plane of the temperature-sensitive hydrogel is not inclined, which indicates that the temperature-sensitive hydrogel can form gel from liquid on the surfaces of skin and mucous membrane, has good gelatinization, and is convenient for attachment and filling. Therefore, the temperature-sensitive hydrogel provided by the invention is still clear and transparent in appearance after being placed for 18 months, and shows sol-gel phase transition behavior at body temperature, and is good in stability.
The in vitro cumulative erosion rate curve (a) and the cumulative release rate curve (B) of the geniposide of the example of fig. 2.
FIG. 3 example shows the evaluation of bacteriostatic effects on E.coli (A), staphylococcus aureus (B), candida albicans (C), pseudomonas aeruginosa (D), streptococcus pneumoniae (E) and Streptococcus pyogenes (F).
Figure 4 example is a recovery graph of rat oral mucosal ulcers.
The example of figure 5 is a graph of percent recovery of oral mucosal ulcer area in rats. * P <0.05 and p <0.0001 compared to the negative control group.
Fig. 6 example 7 results of biocompatibility on rat lingual mucosa.
FIG. 7 is a physical diagram (A) of a white flocculent precipitate produced by directly mixing and contacting chlorhexidine with poloxamer 407 and hyaluronic acid, and a precipitate (B) produced by salting-out effect of NaCl and chlorhexidine dissolved in a poloxamer 407 solution.
Detailed Description
The following description of the present invention will be made clearly and fully, and it is apparent that the embodiments described are some, but not all, of the embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The invention provides poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid, which comprises the following raw material components of hyaluronic acid, chlorhexidine, gardenin, poloxamer 407 and NaCl. Wherein the hyaluronic acid accounts for 0.1 to 0.8 percent, chlorhexidine accounts for 0.05 to 0.1 percent, geniposide accounts for 0.5 percent, poloxamer 407 accounts for 14.0 to 15.0 percent, and NaCl accounts for 0.05 to 0.2 percent. The raw material components are all in weight percentage. In the context of the present invention, the percentages of the raw material components are weight percentages unless otherwise specified.
The invention also provides a specific embodiment of poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid, wherein the hyaluronic acid is 0.5%, the chlorhexidine is 0.1%, the gardenin is 0.5%, the poloxamer 407 is 15.0%, and the NaCl is 0.1-0.2%.
The invention further provides poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid, which comprises the following raw material components, by weight, 0.1-0.8% of hyaluronic acid, 0.05-0.1% of chlorhexidine, 0.5% of gardenin, 14.0-15.0% of poloxamer 407, 0.05-0.2% of NaCl, and the balance of water.
The invention further provides another specific embodiment of poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid, wherein the hyaluronic acid is 0.5%, the chlorhexidine is 0.1%, the gardenin is 0.5%, the poloxamer 407 is 15.0%, and the NaCl is 0.1-0.2%.
The invention also provides a preparation method of the poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid, which comprises the following steps:
step 1): adding chlorhexidine into water for fully dissolving, adding poloxamer 407, standing, and standing at 4 ℃ overnight to fully swell and dissolve;
step 2): adding NaCl into water for dissolution, uniformly scattering hyaluronic acid on the surface of the NaCl, and standing the NaCl to enable the NaCl to be fully swelled and dissolved at room temperature;
step 3): mixing the solutions obtained in the step 1) and the step 2), uniformly stirring, adding the geniposide, and fully stirring and dissolving to obtain the temperature-sensitive hydrogel.
In a specific embodiment provided by the invention, the chlorhexidine is one of chlorhexidine acetate, chlorhexidine gluconate and chlorhexidine hydrochloride.
The invention provides application of the temperature-sensitive hydrogel in preparation of antibacterial anti-inflammatory drugs.
The invention also provides application of the temperature-sensitive hydrogel in preparing a medicament for treating and repairing skin and mucous membrane injury.
The technical scheme of the invention is described in detail below with reference to specific embodiments.
Experimental example 1
Preparation and characterization of poloxamer 407 temperature-sensitive gel and hyaluronic acid modified poloxamer 407 temperature-sensitive gel, and prescription and characterization results are shown in table 1.
The preparation method comprises the following steps:
prepared according to the recipe in Table 1 using the cold solution method.
Preparation of poloxamer 407 temperature-sensitive gel: 100g of poloxamer 407 temperature-sensitive gel is prepared according to the concentration shown in the table 1, and the method comprises the steps of weighing corresponding amounts of poloxamer 407 and deionized water according to the prescription concentration, placing the poloxamer 407 and deionized water in a 100mL beaker, then placing the mixture in a refrigerator at 4 ℃ for full swelling and dissolution, and stirring the mixture to obtain the poloxamer 407 temperature-sensitive gel.
Preparation of hyaluronic acid modified poloxamer 407 temperature-sensitive gel: the method for preparing the poloxamer 407 temperature-sensitive gel modified by the hyaluronic acid comprises the steps of weighing corresponding amounts of poloxamer 407, hyaluronic acid and deionized water according to a prescription, fully swelling and dissolving the hyaluronic acid in 100mL beaker by using deionized water, then adding the poloxamer 407, fully dissolving in a refrigerator at 4 ℃, and stirring to obtain the poloxamer 407 temperature-sensitive gel modified by the hyaluronic acid.
The characterization method comprises the following steps:
(1) Determination of the gel temperature: the test tube inversion method is adopted. 2mL of temperature-sensitive gel solution is filled in a test tube with scales, and a precise thermometer with the precision of 0.1 ℃ is inserted in a constant-temperature water bath kettle. The bulb of the thermometer was completely submerged below the water surface and the gel level was 2cm below the water bath level. Setting the initial temperature of the water bath kettle to 25 ℃ and the temperature rising speed to 0.5 ℃ and min -1 . Taking out the test tube at the temperature of 0.5 ℃ for overturning, observing the gelation condition until the solution solidifies instantly, namely, when no flow is detected within 30 seconds after the test tube is inverted, reading the scale value of the thermometer, wherein the temperature is the gelation temperature, measuring each sample for 3 times in parallel, and taking the average value of the results.
(2) Determination of gel time: taking 2mL of temperature-sensitive gel, placing the temperature-sensitive gel in a 10mL test tube, preheating a sample to 25 ℃, placing the sample in a constant-temperature water bath at 37 ℃, immediately timing by using a stopwatch, and taking the time required when the sample is changed from free flowing liquid to non-flowable semisolid, namely the gelation time. Samples were averaged in triplicate.
(3) Determination of viscosity: 100mL of temperature-sensitive gel solution is placed in a clean and transparent beaker, the clean and transparent beaker is placed in a constant-temperature water bath kettle at 37 ℃ to gel for half an hour to enable the gel to gel completely, the beaker with the gel is kept in the constant-temperature water bath kettle at 37 ℃, the viscosity is measured by a Brookfield rotary rheometer at the speed of a No. 64 rotor and 1.0rpm, and the average value is obtained by three parallel measurement.
(4) Measurement of gel strength: gel strength, i.e., viscosity of the gel at physiological temperature, the present invention quantifies gel strength with a weight of 70g for a time(s) required for 1cm of subsidence in the prepared gel. 30g of the gel was placed in a 50mL measuring cylinder and gelled in a thermostat at 37 ℃. The weight was infiltrated into the gel and the time required for the weight to sink 1cm was recorded. Samples were averaged in triplicate.
Table 1 physicochemical characterization of hyaluronic acid modified poloxamer 407 solution
Note that: "-" indicates that no gel is formed, "600+" indicates that the viscosity value exceeds the measurement range, and "900+" indicates that the intensity value exceeds 900s.
Results:
experiments show that when the concentration of poloxamer 407 is more than 14%, temperature-sensitive gel can be formed, and in order to reduce the dosage of poloxamer 407 and avoid cytotoxicity possibly generated by concentration increase, the poloxamer 407 concentration is selected to be in the range of 14-15% for experiments. As can be seen from table 1:
(1) As the concentration of poloxamer 407 increases, the gelation temperature and gelation time gradually decrease, and the viscosity and strength of the formed gel gradually increase.
(2) When the concentration of hyaluronic acid is less than 0.5%, gel cannot be formed if the concentration of poloxamer 407 is less than 14.2%; when the concentration of hyaluronic acid is more than 0.5%, the viscosity of the hyaluronic acid/poloxamer 407 solution is too high to be injected easily. The concentration of hyaluronic acid was chosen to be 0.5% for this purpose, and the poloxamer 407 concentration was further defined as between 14.2 and 15%.
(3) The gel temperature and gel time of the 0.5% hyaluronic acid modified poloxamer 407 gel is reduced compared with unmodified gel, but the viscosity and strength are increased sharply, and the measurement range is exceeded.
Experimental example 2
In order to reduce the strength and viscosity of the hyaluronic acid modified poloxamer 407 temperature-sensitive gel, the electro-adhesion effect of the NaCl barrier part was selected, and the preparation method of the NaCl-regulated hyaluronic acid/poloxamer 407 temperature-sensitive gel is as follows, and the prescription is shown in Table 2.
The preparation method comprises the following steps:
100g of each of the series of solutions having a hyaluronic acid concentration of 0.5%, a NaCl concentration of 0.05-0.2% and a poloxamer 407 concentration of 14-15% were prepared according to the prescription of Table 2. Weighing corresponding amounts of poloxamer 407, hyaluronic acid and deionized water according to a prescription, fully swelling and dissolving the hyaluronic acid in a 100mL beaker by using deionized water, then adding the poloxamer 407, standing in a refrigerator at 4 ℃ for overnight, fully swelling and dissolving, uniformly stirring, then adding the prescribed amount of NaCl, and uniformly stirring to obtain the aqueous emulsion.
The characterization method comprises the following steps: the results are shown in Table 2, similar to Experimental example 1.
Results:
as the NaCl concentration increases, the concentration of poloxamer 407 required to form a gel gradually increases.
In the prescription capable of forming gel, the gelation temperature is slightly increased along with the increase of NaCl concentration, and the maximum temperature is 29.8 ℃, so that the gel is suitable for being used at body temperature; the gel time is gradually increased, and the maximum time is 209.53s, which is not more than 4min; the viscosity and strength of the gel are significantly reduced. Therefore, naCl can effectively regulate the viscosity and strength of the hyaluronic acid modified poloxamer 407 temperature-sensitive gel, so that the hyaluronic acid modified poloxamer 407 temperature-sensitive gel is easy to attach to skin and mucous membrane.
(3) In order to form temperature-sensitive gel, and the formed gel has proper physicochemical properties, so that the medication comfort and the compliance of patients are improved, the dosage of NaCl is selected to be 0.15-0.2%, and the dosage of poloxamer 407 is selected to be 14.8-15.0%.
TABLE 2 Effect of different concentrations of NaCl on poloxamer 407/hyaluronic acid composite hydrogel parameters
Note that: "-" means that the solution is not temperature sensitive and "900+" means that the intensity value exceeds 900s.
Examples 1 to 10
Poloxamer 407 temperature-sensitive hydrogel containing hyaluronic acid/chlorhexidine/geniposide
100g of the temperature-sensitive hydrogel solutions of examples 1 to 10 were prepared according to the formulations shown in Table 3, respectively.
(1) Poloxamer 407, chlorhexidine, geniposide, naCl and hyaluronic acid are respectively and precisely weighed according to the prescription composition.
(2) Deionized water was weighed according to the prescription composition and placed in two 100mL beakers, designated beaker 1 and beaker 2, in approximately a 1:1 ratio.
(3) The weighed chlorhexidine was added to beaker 1, stirred until completely dissolved, then the weighed poloxamer 407 was added, and then the mixture was placed in a refrigerator at 4 ℃ for refrigeration overnight to allow the mixture to slowly swell and dissolve to give a clear and transparent solution.
(4) In examples 1-5, the weighed hyaluronic acid powder was uniformly sprinkled on the deionized water level of beaker 2, and swollen and dissolved at room temperature until a clear and transparent solution was obtained. In examples 6-10, the weighed NaCl was completely dissolved in deionized water in beaker 2, and then the weighed hyaluronic acid powder was uniformly sprinkled on the liquid surface thereof, and swelled and dissolved at room temperature until a clear and transparent solution was obtained.
(5) Mixing the two solutions in the beaker 1 and the beaker 2, adding the weighed geniposide after stirring uniformly, and stirring until the geniposide is fully dissolved to obtain the transparent and clear poloxamer 407 temperature-sensitive hydrogel solution containing hyaluronic acid/chlorhexidine/geniposide, which has no lumps and is uniformly dispersed.
Table 3 prescription composition of examples 1-10
Note that: in the table "-" indicates that the component is absent, and the addition step of the component is omitted in the corresponding preparation method.
Evaluation example 1
In vitro erosion and drug Release test of examples 1-7
The experimental method comprises the following steps: and (3) determining the gel erosion rate and the drug release rate by adopting a membraneless dissolution method.
(1) The corrosion rate measurement method takes 7 test tubes of 10mL respectively numbered as tubes 1,2,3,4,5,6,7, and then respectively weighing, and the mass is recorded as Wi-0, wherein i corresponds to the number of the test tube. After 1mL of the gel solutions of examples 1-7, respectively, were placed in the corresponding numbered test tubes, they were rapidly weighed and the mass was recorded as Wi-1, where i is also the number corresponding to the test tube. Then, 7 test tubes are placed in a constant temperature water bath kettle at 37 ℃ for 0.5h, 3mL of phosphate buffer solution with the pH of 6.68 preheated to the same temperature is gently added one by one along the walls of the test tubes after the test tubes are completely gelled, and the adding process is slow to keep the gel liquid level from damaging. Then 7 test tubes are put into a constant temperature water bath oscillator at 37 ℃ and 50r/min for oscillation, the upper solution is poured out at 0.5, 1,2, 4, 6, 8, 10, 12 and 13h respectively, the medium is released from the surface of the test tube after the test tube is sucked by filter paper, the test tube is rapidly and precisely weighed, the quality is recorded as Wi-j, i corresponds to the serial number of the test tube, and j corresponds to the measurement time. After weighing, each tube was immediately replenished with isothermal 3mL of phosphate buffer, and the shaking was continued at constant temperature, and the above operation was repeated for 13 hours. The cumulative erosion rate (Rc%) for each test site was calculated according to equation (1). Each example was repeated 3 times and the results averaged and the erosion curve is shown in figure 2-a.
(2) Method for measuring drug release the drug release was studied in examples 4,5,6 and 7 while measuring the dissolution rate in vitro. 1mL of the release medium poured out at each time point of examples 4 to 7 was precisely aspirated, placed in a 5mL EP tube, 1mL of phosphate buffer solution pH 6.68 was added, sonicated for 20min, the concentration of geniposide in the sample was measured by HPLC method and the cumulative drug release rate was calculated according to formulas 2 and 3. Each sample was measured 3 times in parallel and the results averaged and the cumulative release curve is shown in figure 2-B.
Wherein n is the sampling time point, X n Drug release amount at time n, C n For the drug concentration measured at time n, V n For a sample volume (1 mL), i is any sampling point before time n.
Wherein F is the cumulative release rate, M is the amount of drug contained in 1mL of gel, and the unit is mg.
Results:
as can be seen from fig. 2-a, the gel erosion rate gradually decreased as the concentration of poloxamer 407 increased from 14.2% to 15.0% in examples 1-5. The reason is probably that as the concentration of poloxamer 407 increases, the number and size of micelles increases within the gel-crosslinked three-dimensional structure, causing the number and size of water channels to decrease, resulting in a decrease in the gel erosion rate.
Examples 6 and 7 are examples 4 and 5, respectively, with 0.15% and 0.2% NaCl, and it is apparent that the addition of NaCl significantly increases the erosion rate.
(3) The literature shows that the faster the gel erodes, the faster the drug is released. By comparing FIG. 2-A with FIG. 2-B, it can be obtained that the addition of NaCl increases the dissolution rate of the gel and simultaneously increases the release rate of the geniposide from the gel, which is consistent with the results of the literature. The reason is that the high hydrophilicity of NaCl can easily diffuse out of the gel water channel, and the precipitated sodium salt can leave pores in the gel, so that external liquid can more easily enter the gel matrix, and the diffusion and corrosion speed of the gel can be increased.
(4) The drug release data fitting of example 7 shows that the release of the geniposide accords with the zero order kinetics law, which indicates that the drug is released continuously at a constant rate, avoids any initial burst release (peak Gu Xianxiang), is beneficial to reducing toxic and side effects and improving the curative effect of the drug.
Evaluation example 2
Examination of bacteriostatic Effect of examples 7 and 8
The experimental method comprises the following steps: taking deionized water as a negative control, taking commercial chlorhexidine (0.05-0.1%) mouthwash as a positive control, and examining the antibacterial effect of examples 7 and 8 on escherichia coli, staphylococcus aureus, candida albicans, pseudomonas aeruginosa, streptococcus pneumoniae and streptococcus pyogenes.
1 colony was collected from LB agar plates of E.coli, staphylococcus aureus, candida albicans, pseudomonas aeruginosa, streptococcus pneumoniae and Streptococcus pyogenes, respectively, and placed in 15mL culture tubes containing LB liquid medium, respectively. The culture tube was then placed in an incubator at 37℃and 200rpm for 12 hours. Shake flasks containing 150mL of LB liquid medium were sterilized at 120 ℃ for 40min and cooled. 2mL of the bacterial liquid in the liquid culture tube is sucked into a shake flask, 3mL of a sample to be tested is added at the same time, and the mixture is placed in an incubator at 37 ℃ and 200 rpm. 1mL of the sample was sampled every 1 hour, shaken, dropped into a 96-well plate with a 200. Mu.L pipette, and three wells were dropped in parallel, and the Optical Density (OD) at 600nm was measured with a microplate reader, and the result was averaged three times. The bacterial concentration was quantified by OD value, and an OD value of 0.1 corresponds to a bacterial concentration of 108 pieces/cm 3. The bacterial growth curve was plotted with time t on the abscissa and OD on the ordinate for 11 hours of continuous sampling, and the results are shown in fig. 3.
Analysis of results:
(1) From fig. 3A-F, the OD values of the control group were significantly higher than those of the positive control group and the example group during the measurement time, indicating that the positive control group and the example group had different degrees of inhibition on 6 bacteria.
(2) The OD values of the positive control groups in fig. 3A-D were significantly higher than those of the example groups, demonstrating that the antibacterial effect of examples 7, 8 on escherichia coli, staphylococcus aureus, candida albicans, pseudomonas aeruginosa was significantly better than that of the commercial chlorhexidine mouthwash.
(3) In FIGS. 3A-F, the OD value of example 7 was almost unchanged within 0-11h, maintaining a small level, demonstrating that example 7 had good inhibition against 6 bacteria. Example 8 only in figure 3D the OD value increases with time, demonstrating that the bacteriostatic effect of chlorhexidine on pseudomonas aeruginosa is affected by concentration, with poor effect at low concentrations; but increased to a significantly lower extent than the positive control, indicating that it still has a level of inhibition of pseudomonas aeruginosa growth that is superior to the commercial mouthwash.
(4) In fig. 3E-F, the bacterial growth rate curves of examples 7, 8 and the positive control almost coincide, the OD value is very low and hardly changes with time, indicating that the bacteriostatic effect of the low concentration chlorhexidine gel (example 8) and the commercial mouthwash on streptococcus pneumoniae and streptococcus pyogenes is identical to that of example 7 at high concentration, and the effect is very good.
Evaluation example 3
Examples 7, 9 and 10 recovery effects on rat oral mucosal ulcers
The method comprises the following steps: healthy male Wistar albino rats were randomly divided into 15 groups of 3 Normal control (Normal), negative control (NegCon), example 7, example 9 and example 10. Normal control rats were not induced by oral ulcer, no treatment/administration was performed, and other rats were induced by oral ulcer. Rats in the negative control group were not treated, and the other groups were given 0.1 g/time of the corresponding example temperature sensitive gel, respectively. Rats were anesthetized by intraperitoneal injection of chloral hydrate (0.4 mL of chloral hydrate at a concentration of 10% per 100 grams of rats) to allow the oral mucosal ulcer induction procedure to proceed smoothly. 20. Mu.L of 70% acetic acid was dipped in 5mm length and width filter paper and placed in the middle of the back of the rat tongue for 2min to induce oral mucosal ulcers. On day 1 of ulcer formation, the maximum ulcer area (ulcer area was 100%) was determined. Topical administration was started 1 day from the time of ulcer development, 1 time/d, and continued for 7 days. Record every other day by taking picturesComputer software calculates the recovery rate of the ulcer area of the rat by circularly measuring the ulcer area on the ulcer edge using formula 4, and the results are shown in fig. 4 and 5.
Analysis of results:
as can be seen from FIGS. 4 and 5, after one week of treatment, the ulcers in each group were significantly improved.
Example 10 has an ulcer area recovery of about 76% and higher than that of example 9, indicating that hyaluronic acid has a promoting effect on injury repair.
The mucosa treated in example 7 had a most normal appearance (fig. 4) and a highest recovery rate of ulcer area of about 86%, indicating that the antibacterial agent chlorhexidine and the anti-inflammatory agent geniposide play a role in oral mucosa ulcer recovery.
Evaluation example 4
Example 7 evaluation of biocompatibility
The experimental method comprises the following steps: 6 healthy male Wistar albino rats were divided into a normal control group and a test group, each group having 3 animals. The normal control group did not undergo any treatment. The test group was given 0.1 g/time of the gel of example 7 on the mid-lingual side of the rats daily for 7 consecutive days. Rat tongue samples were stored in paraformaldehyde solution for HE staining and the results are shown in fig. 6.
Analysis of results: FIGS. 6-A and 6-B are, respectively, images of normal tongue mucosa tissue and tongue mucosa tissue taken 7 days after administration of the gel of example 7, which are very similar, showing healthy skeletal muscle fibers, no significant inflammation or necrosis, demonstrating that the hydrogel set of example 7 has a high degree of biocompatibility.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.
Claims (5)
1. The preparation method of the poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenia glycoside/hyaluronic acid is characterized by comprising the following steps of:
step 1): adding chlorhexidine into water for fully dissolving, adding poloxamer 407, standing, and standing at 4 ℃ overnight to fully swell and dissolve;
step 2): adding NaCl into water for dissolution, uniformly scattering hyaluronic acid on the surface of the NaCl, and standing the NaCl to enable the NaCl to be fully swelled and dissolved at room temperature;
step 3): mixing the solutions obtained in the step 1) and the step 2), uniformly stirring, adding geniposide, and fully stirring and dissolving to obtain temperature-sensitive hydrogel; the hyaluronic acid accounts for 0.5 percent, the chlorhexidine accounts for 0.1 percent, the geniposide accounts for 0.5 percent, the poloxamer 407 accounts for 14.8 to 15.0 percent, and the NaCl accounts for 0.15 to 0.2 percent.
2. The method for preparing the poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid according to claim 1, wherein the chlorhexidine is one of chlorhexidine acetate, chlorhexidine gluconate and chlorhexidine hydrochloride.
3. A temperature-sensitive hydrogel prepared by a preparation method of poloxamer 407 antibacterial anti-inflammatory temperature-sensitive hydrogel containing chlorhexidine/gardenin/hyaluronic acid according to claim 2.
4. Use of the temperature-sensitive hydrogel of claim 3 in the preparation of antibacterial drugs.
5. Use of the temperature-sensitive hydrogel of claim 3 in the preparation of a medicament for treating and repairing mucosal injury.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IN183804B (en) * | 1997-10-29 | 2000-04-22 | M S Lekar Pharma Ltd | |
| BR0206336A (en) * | 2002-12-06 | 2004-10-05 | Univ Minas Gerais | Process of Preparation of Slow Release Antimicrobial, Anesthetic, Antifungal and Anti-Inflammatory Pharmaceutical Compositions and Derived Products |
| CN102552974A (en) * | 2012-02-17 | 2012-07-11 | 上海白衣缘生物工程有限公司 | Gel composition for injection filling of skins and preparation method for gel composition |
| CN103860694A (en) * | 2014-02-14 | 2014-06-18 | 四川蜀源博业医药科技有限公司 | Ion-sensitive in situ gel for nasal cavity and preparation method and application thereof |
| CN105168238A (en) * | 2015-08-13 | 2015-12-23 | 南京天纵易康生物科技股份有限公司 | Hyaluronic temperature-sensitive gel and preparation method and application thereof |
| WO2021036058A1 (en) * | 2019-08-28 | 2021-03-04 | 深圳翰宇药业股份有限公司 | Teriparatide sustained release gel injection solution and preparation method therefor |
| CN113521290A (en) * | 2020-04-16 | 2021-10-22 | 正执(上海)医疗器械有限公司 | Pharmaceutical composition, medicinal patch, and preparation method and application thereof |
-
2022
- 2022-10-24 CN CN202211303056.5A patent/CN115634234B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IN183804B (en) * | 1997-10-29 | 2000-04-22 | M S Lekar Pharma Ltd | |
| BR0206336A (en) * | 2002-12-06 | 2004-10-05 | Univ Minas Gerais | Process of Preparation of Slow Release Antimicrobial, Anesthetic, Antifungal and Anti-Inflammatory Pharmaceutical Compositions and Derived Products |
| CN102552974A (en) * | 2012-02-17 | 2012-07-11 | 上海白衣缘生物工程有限公司 | Gel composition for injection filling of skins and preparation method for gel composition |
| CN103860694A (en) * | 2014-02-14 | 2014-06-18 | 四川蜀源博业医药科技有限公司 | Ion-sensitive in situ gel for nasal cavity and preparation method and application thereof |
| CN105168238A (en) * | 2015-08-13 | 2015-12-23 | 南京天纵易康生物科技股份有限公司 | Hyaluronic temperature-sensitive gel and preparation method and application thereof |
| WO2021036058A1 (en) * | 2019-08-28 | 2021-03-04 | 深圳翰宇药业股份有限公司 | Teriparatide sustained release gel injection solution and preparation method therefor |
| CN113521290A (en) * | 2020-04-16 | 2021-10-22 | 正执(上海)医疗器械有限公司 | Pharmaceutical composition, medicinal patch, and preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 快速环境响应多孔水凝胶的制备及性能;刘云海等;《高分子材料科学与工程》;第22卷(第5期);第233页第2.1部分；摘要 * |
| 栀子及栀子苷抗炎作用机制的最新研究进展;万亮琴等;《现代中药研究与实践》;第31卷(第3期);摘要；第81页左栏部分 * |
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